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J Neurosci Methods ; 218(1): 1-8, 2013 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-23660525

RESUMO

A novel, automated system for delivering controlled scratch-induced trauma to brain cells cultured in multi-well plates was created and characterized. The system is equipped with high-throughput imaging and analysis capabilities, enabling quantitative measurements of cell migration. The scratch-area coefficient of variation of the device was between 3.9% and 8.4%, a significant improvement over traditional manual methods, which provided a scratch-area coefficient of variation of between 10.7% and 19.6%. The device's inexpensive imaging and analysis capabilities were comparable to a well-known system, the Discovery-1 (Molecular Devices), with no significant difference found between the two. When used for drug screening, the gap area of Neuro2a cells after 72h was significantly larger in samples containing UO126 (20µM), averaging 0.89mm(2)±0.21mm(2); compared with an average vehicle control gap area of 0.42mm(2)±0.1mm(2). A gradient response could also be detected among samples with increasing UO126 concentrations (0-20µM), due to decreased migration and/or proliferation of cells into the gap over the time period. Our device provides an inexpensive method for delivering a standardized, closely controlled pressure/scratch to brain cells cultured in multi-well plates. The system provides more consistent patterns of scratch-induced trauma to cultured cells when compared to traditional methods. This device is an effective platform for quantifying the injury response of cells, and has applications in testing the effectiveness of drugs on cell migration and proliferation which might potentially treat traumatic brain injury.


Assuntos
Lesões Encefálicas , Ensaios de Triagem em Larga Escala/instrumentação , Neurônios , Animais , Movimento Celular , Células Cultivadas , Camundongos
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